Comparison of exopolysaccharide production by strains of Lactobacillus rhamnosus and Lactobacillus paracasei grown in chemically defined medium and milk

Exopolysaccharide (EPS) production was compared among three strains of lactobacilli. Lactobacillus rhamnosus strain 9595M can be classified among the highest EPS-producing strains of lactic acid bacteria reported to date with a maximum EPS production of 1275 mg L⁻¹. Under controlled pH, no significant differences in the quantity of EPS produced could be detected between carbon source (glucose or lactose) or fermentation temperature (32 or 37°C). In milk, strains ATCC 9595M and R produced more than 280 mg L⁻¹ EPS whereas strain Type V produced less than 80 mg L⁻¹ EPS. Journal of Industrial Microbiology & Biotechnology (2000) 24, 251–255. Received 10 September 1999/ Accepted in revised form 22 December 1999     

Thermal acclimation of locomotor performance in tadpoles and adults of the aquatic frog Xenopus laevis

Among amphibians, the ability to compensate for the effects of temperature on the locomotor system by thermal acclimation has only been reported in larvae of a single species of anuran. All other analyses have examined predominantly terrestrial adult life stages of amphibians and found no evidence of thermal acclimatory capacity. We examined the ability of both tadpoles and adults of the fully aquatic amphibian Xenopus laevis to acclimate their locomotor system to different temperatures. Tadpoles were acclimated to either 12 °C or 30 °C for 4 weeks and their burst swimming performance was assessed at four temperatures between 5 °C and 30 °C. Adult X. laevis were acclimated to either 10 °C or 25 °C for 6 weeks and their burst swimming performance and isolated muscle performance was determined at six temperatures between 5 °C and 30 °C. Maximum swimming performance of cold-acclimated X. laevis tadpoles was greater at cool temperatures and lower at the highest temperature in comparison with the warm-acclimated animals. At the test temperature of 12 °C, maximum swimming velocity of tadpoles acclimated to 12 °C was 38% higher than the 30 °C-acclimation group, while at 30 °C, maximum swimming velocity of the 30 °C-acclimation group was 41% faster than the 12 °C-acclimation group. Maximum swimming performance of adult X. laevis acclimated to 10 °C was also higher at the lower temperatures than the 25 °C acclimated animals, but there was no difference between the treatment groups at higher temperatures. When tested at 10 °C, maximum swimming velocity of the 10 °C-acclimation group was 67% faster than the 25 °C group. Isolated gastrocnemius muscle fibres from adult X. laevis acclimated to 10 °C produced higher relative tetanic tensions and decreased relaxation times at 10 °C in comparison with animals acclimated to 25 °C. This is only the second species of amphibian, and the first adult life stage, reported to have the capacity to thermally acclimate locomotor performance. Accepted: 28 October 1999     

Screening and characterization of bioflocculant produced by isolated Klebsiella sp.

Sixteen strains of polymer-producing bacteria were isolated from the activated sludge samples taken from two seafood processing plants in Southern Thailand. Their culture broths possessed the ability to flocculate kaolin suspension in the presence of 1% CaCl₂. Based on the flocculating activity, the strain S11 was selected and identified to be a Klebsiella sp. using the partial 16S rRNA sequencing method. The growth of the isolated Klebsiella sp. was maximal (1.026 g l⁻¹ dry cell mass) after 1 day cultivation while the highest polymer yield (0.973 g l⁻¹) was achieved after 5 days cultivation. The flocculating activity of the culture broth, however, was highest after 2 days cultivation. The polymer was identified to be an acidic polysaccharide containing neutral sugar and uronic acid as its major and minor components, respectively. Results on the properties of the partially purified polysaccharide from Klebsiella sp. S11 revealed that it consisted of galactose, glucose and mannose in an approximate ratio of 5:2:1. It was soluble in acidic or basic solutions but not in organic solvents. Its molecular mass was greater than 2 × 10⁶ Da. Infrared spectra showed the presence of hydroxyl, carboxyl and methoxyl groups in its molecules. Differential scanning calorimetry of the polysaccharide indicated the crystalline melting point (T _m) at 314 °C. The optimum dosage of polysaccharide to give the highest flocculating activity was 15 mg l⁻¹ in the presence of 1% CaCl₂. Received: 8 February 1999 / Received last revision: 4 June 1999 / Accepted: 4 June 1999     

cAMP-dependent protein kinase from brown adipose tissue: temperature effects on kinetic properties and enzyme role in hibernating ground squirrels

Arousal from hibernation requires thermogenesis in brown adipose tissue, a process that is stimulated by β-adrenergic signals, leading to a rise in intracellular 3′,5′-cyclic adenosine monophosphate AMP (cAMP) and activating cAMP-dependent protein kinase A (PKA) to phosphorylate a suite of target proteins and activate lipolysis and uncoupled respiration. To determine whether specific adaptations (perhaps temperature-dependent) facilitate PKA kinetic properties or protein-phosphorylating ability, the catalytic subunit of PKA (PKAc) from interscapular brown adipose of the ground squirrel Spermophilus richardsonii, was purified (final specific activity = 279 nmol phosphate transferred per min per mg protein) and characterized. Physical properties of PKAc included a molecular weight of 41 kDa and an isoelectric point of 7.8 ± 0.08. A change in assay temperature from a euthermic value (37 °C) to one typical of hibernating body temperature (5 °C) had numerous significant effects on ground squirrel PKAc including: (a) pH optimum rose from 6.8 at 37 °C to 8.7 at 5 °C, (b) K_m values at 37 °C for Mg.ATP (49.2±3.4 M) and for two phosphate acceptors, Kemptide (50.0±5.5 M) and Histone IIA (0.41 ± 0.05 mg/ml) decreased by 53%, 80% and 51%, respectively, at 5 °C, and (c) inhibition by KCl, NaCl and NH₄Cl was reduced. However, temperature change had little or no effect on K_m values of rabbit PKAc, suggesting a specific positive thermal modulation of the hibernator enzyme. Arrhenius plots also differed for the two enzymes; ground squirrel PKAc showed a break in the Arrhenius relationship at 9 °C and activation energies that were 29.1 ± 1.0 kJ/mol for temperatures >9 °C and 2.3-fold higher at 68.1 ± 2.1 kJ/mol for temperatures <9 °C, whereas the rabbit enzyme showed a breakpoint at 17 °C with a 13-fold higher activation energy over the lower temperature range. However, fluorescence analysis of PKAc in the absence of substrates, showed a linear change in fluorescence intensity and wavelength of maximal fluorescence over the entire temperature range; this suggested that the protein conformational change indicated by the break in the Arrhenius plot was substrate-related. Temperature change also affected the Hill coefficient for cAMP dissociation of the ground squirrel PKA holoenzyme which rose from 1.12 ± 0.18 at 37 °C to 2.19 ± 0.07 at 5 °C, making the release of catalytic subunits at low temperature much more responsive to small changes in cAMP levels. Analysis of PKAc function via in vitro incubations of extracts of ground squirrel brown adipose with ³²P-ATP + cAMP in the presence versus absence of a PKA inhibitor, also revealed major differences in the patterns of phosphoproteins, both between euthermic and hibernating animals as well as between 37 and 5 °C incubation temperatures; this suggests that there are both different targets of PKAc phosphorylation in the hibernating animal and that temperature affects the capacity of PKAc to phosphorylate different targets. Both of these observations, plus the species-specific and temperature-dependent changes in ground squirrel PKAc kinetic properties, suggest differential control of the enzyme in vivo at euthermic versus hibernating body temperatures in a manner that would facilitate a rapid and large activation of the enzyme during arousal from torpor. Accepted: 10 July 1998     

Production of poly-β-hydroxybutyrate (PHB) by <Emphasis Type="Italic">Alcaligenes latus</Emphasis> from maple sap

Maple sap, an abundant natural product especially in Canada, is rich in sucrose and thus may represent an ideal renewable feedstock for the production of a wide variety of value-added products. In the present study, maple sap or sucrose was employed as a carbon source to Alcaligenes latus for the production of poly-β-hydroxybutyrate (PHB). In shake flasks, the biomass obtained from both the sap and sucrose were 4.4 ± 0.5 and 2.9 ± 0.3 g/L, and the PHB contents were 77.6 ± 1.5 and 74.1 ± 2.0%, respectively. Subsequent batch fermentation (10 L sap) resulted in the formation of 4.2 ± 0.3 g/L biomass and a PHB content of 77.0 ± 2.6%. The number average molecular weights of the PHB produced by A. latus from maple sap and pure sucrose media were 300 ± 66 × 10³ and 313 ± 104 × 10³ g/mol, respectively. Near-infrared, ¹H magnetic resonance imaging (MRI), and ¹³C-MRI spectra of the microbially produced PHB completely matched those obtained with a reference material of poly[(R)-3-hydroxybutyric acid]. The polymer was found to be optically active with [α]²⁵ _D equaled to −7.87 in chloroform. The melting point (177.0°C) and enthalpy of fusion (77.2 J/g) of the polymer were also in line with those reported, i.e., 177°C and 81 J/g, respectively.     

A neutral carrier-based liquid membrane microelectrode for divalent putrescine cations

A new ion-selective liquid membrane microelectrode, based on the neutral carrier 1,1′-bis(2,3-naphtho-18-crown-6), is described that shows the dependence of EMF on the activity of divalent putrescine cations a _Put, with the linear slope s _Put = 26 ± 3 mV/decade (mean ± SD, N = 18), in the range 10⁻⁴–10⁻¹ M at 25 ± 1 °C. Values of potentiometric putrescine cation selectivity coefficients of logK ^Pot _{Put j} (mean ± SD, N) are obtained by the separate solution method for the ions K⁺ (1.0 ± 0.4, 10), Na⁺ (−1.2 ± 0.4, 8), Ca²⁺ (−2.3 ± 0.5, 10) and Mg²⁺ (−2.5 ± 0.5, 7). The microelectrode can be applied for the direct analysis of the activities of free divalent putrescine cations in the range 5 × 10⁻⁴ to 10⁻¹ M in an extracellular ionic environment. Established analytical methods, e.g. high performance liquid chromatography, determine the total concentration of the derivatives of free and bound putrescine. Received: 20 December 1998 / Revised version: 7 May 1999 / Accepted: 27 May 1999     

Thermal ecology of the Australian agamid Pogona barbata

This study compares the thermal ecology of male bearded dragon lizards (Pogona barbata) from south-east Queensland across two seasons: summer (1994–1995) and autumn (1995). Seasonal patterns of body temperature (T _b) were explored in terms of changes in the physical properties of the thermal environment and thermoregulatory effort. To quantify thermoregulatory effort, we compared behavioral and physiological variables recorded for observed lizards with those estimated for a thermoconforming lizard. The study lizards' field T _bs varied seasonally (summer: grand daily mean (GDM) 34.6 ± 0.6°C, autumn: GDM 27.5 ± 0.3°C) as did maximum and minimum available operative temperatures (summer: GDM T _max 42.1 ± 1.7°C, T _min 32.2 ± 1.0°C, autumn: GDM T _max 31.7 ± 1.2°C, T _min 26.4 ± 0.5°C). Interestingly, the range of temperatures that lizards selected in a gradient (selected range) did not change seasonally. However, P. barbata thermoregulated more extensively and more accurately in summer than in autumn; lizards generally displayed behaviors affecting heat load nonrandomly in summer and randomly in autumn, leading to the GDM of the mean deviations of lizards' field T _bs from their selected ranges being only 2.1 ± 0.5°C in summer, compared to 4.4 ± 0.5°C in autumn. This seasonal difference was not a consequence of different heat availability in the two seasons, because the seasonally available ranges of operative temperatures rarely precluded lizards from attaining field T _bs within their selected range, should that have been the goal. Rather, thermal microhabitat distribution and social behavior appear to have had an important influence on seasonal levels of thermoregulatory effort. Received: 28 April 1997 / Accepted: 29 December 1997     

Thermal acclimation of locomotor performance in tadpoles of the frog Limnodynastes peronii

Previous analyses of thermal acclimation of locomotor performance in amphibians have only examined the adult life history stage and indicate that the locomotor system is unable to undergo acclimatory changes to temperature. In this study, we examined the ability of tadpoles of the striped marsh frog (Limnodynastes peronii) to acclimate their locomotor system by exposing them to either 10 °C or 24 °C for 6 weeks and testing their burst swimming performance at 10, 24, and 34 °C. At the test temperature of 10 °C, maximum velocity (U_max) of the 10 °C-acclimated tadpoles was 47% greater and maximum acceleration (A_max) 53% greater than the 24 °C-acclimated animals. At 24 °C, U_max was 16% greater in the 10 °C-acclimation group, while there was no significant difference in A_max or the time taken to reach U_max (T-U_max). At 34 °C, there was no difference between the acclimation groups in either U_max or A_max, however T-U_max was 36% faster in the 24 °C-acclimation group. This is the first study to report an amphibian (larva or adult) possessing the capacity to compensate for cool temperatures by thermal acclimation of locomotor performance. To determine whether acclimation period affected the magnitude of the acclimatory response, we also acclimated tadpoles of L. peronii to 10 °C for 8 months and compared their swimming performance with tadpoles acclimated to 10 °C for 6 weeks. At the test temperatures of 24 °C and 34 °C, U_max and A_max were significantly slower in the tadpoles acclimated to 10 °C for 8 months. At 10 °C, T-U_max was 40% faster in the 8-month group, while there were no differences in either U_max or A_max. Although locomotor performance was enhanced at 10 °C by a longer acclimation period, this was at the expense of performance at higher temperatures. Accepted: 25 June 1999     

Quercetin in Lyotropic Liquid Crystalline Formulations: Physical, Chemical and Functional Stability

The purpose of this study was to develop a lyotropic liquid crystalline formulation using the emulsifier vitamin E TPGS and evaluate its behavior after incorporation of a flavonoid, quercetin. The physical (macro and microscopic), chemical (determination of quercetin content by the HPLC method) and functional (determination of quercetin antioxidant activity by DPPH^• assay) stability of the lamellar liquid crystalline formulation containing flavonoid was evaluated when stored at 4 ± 2 °C; 30 ± 2 °C/70 ± 5% RH (relative humidity) and 40 ± 2 °C/70 ± 5% RH during 12 months. The lamellar liquid crystalline structure of the formulation was maintained during the experiment, however chemical and functional stability results showed a great influence of the storage period in all conditions tested. A significant decrease in quercetin content (approximately 40%) was detected during the first month of storage and a similar significant loss in antioxidant activity was detected after 6 months. The remaining flavonoid content was unchanged during the final 6 months of the experimental period. The results suggest possible interactions between quercetin and the liquid crystalline formulation, which could inhibit or reduce the quercetin activity incorporated in the system. In conclusion, the present study demonstrated that incorporation of quercetin (1%) did not affect the liquid crystalline structure composed of vitamin E TPGS/IPM/PG–H₂O (1:1) at 63.75/21.25/15 (w/w/w). Nevertheless, of the total quercetin incorporated in the system only 60% was free to act as an antioxidant.     

Batch and fed-batch bioreactor cultivations of a Thermus species with thermophilic BTEX-degrading activity

The thermophilic bacterium, Thermus species ATCC 27978, which is capable of aerobically degrading benzene, toluene, ethylbenzene, and the xylenes (BTEX), was cultured in 5-1 fermentors on a Castenholz salts-tryptone medium. This bacterium can be cultivated more conveniently at 45 °C, a temperature substantially lower than its optimal growth temperature (approx. 60 °C). Yet, the washed harvested cells from such cultures display the same initial BTEX-degrading activity as those when Thermus sp. is grown at its higher optimal temperature. Two bioreactor cultivation modes, batch and fed batch, were investigated. More biomass and more BTEX-degrading activity (assayed at 60 °C) were generated in fed-batch cultures than in the growth-limited batch cultures. The former yielded a biomass concentration of 2.5 g dry cell weight (DCW) l⁻¹ and whole-cell degrading specific activities of 7.6 ± 1.3, 10.1 ± 1.9, 9.8 ± 2.1, 2.3 ± 0.5, and 4.6 ± 0.9 nmol degraded (mg DCW)⁻¹ min⁻¹ for benzene, toluene, ethylbenzene, m-xylene, and the o- plus p-xylenes (unresolved mixture), respectively. Although the formation of cellular BTEX-degrading activity is growth-associated, a slow to moderate specific growth rate of 0.02–0.07 h⁻¹ favors the production of BTEX-degrading activity, while a high growth rate, of the order of 0.16 h⁻¹, is detrimental to its production. The washed harvested Thermus sp. cells were capable of degrading BTEX over a broad range of thermophilic incubation temperatures, 45–77 °C. Received: 28 June 1996 / Received revision: 31 December 1996 / Accepted: 31 January 1997     

Comparison of separate hydrolysis and fermentation and simultaneous saccharification and fermentation processes for ethanol production from wheat straw by recombinant <Emphasis Type="Italic">Escherichia coli</Emphasis> strain FBR5

Ethanol production by recombinant Escherichia coli strain FBR5 from dilute acid pretreated wheat straw (WS) by separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF) was studied. The yield of total sugars from dilute acid (0.5% H₂SO₄) pretreated (160 °C, 10 min) and enzymatically saccharified (pH 5.0, 45 °C, 72 h) WS (86 g/l) was 50.0 ± 1.4 g/l. The hydrolyzate contained 1,184 ± 19 mg furfural and 161 ± 1 mg hydroxymethyl furfural per liter. The recombinant E. coli FBR5 could not grow at all at pH controlled at 4.5 to 6.5 in the non-abated wheat straw hydrolyzate (WSH) at 35 °C. However, it produced 21.9 ± 0.3 g ethanol from non-abated WSH (total sugars, 44.1 ± 0.4 g/l) in 90 h including the lag time of 24 h at controlled pH 7.0 and 35 °C. The bioabatement of WS was performed by growing Coniochaeta ligniaria NRRL 30616 in the liquid portion of the pretreated WS aerobically at pH 6.5 and 30 °C for 15 h. The bacterium produced 21.6 ± 0.5 g ethanol per liter in 40 h from the bioabated enzymatically saccharified WSH (total sugars, 44.1 ± 0.4 g) at pH 6.0. It produced 24.9 ± 0.3 g ethanol in 96 h and 26.7 ± 0.0 g ethanol in 72 h per liter from bioabated WSH by batch SSF and fed-batch SSF, respectively. SSF offered a distinct advantage over SHF with respect to reducing total time required to produce ethanol from the bioabated WS. Also, fed-batch SSF performed better than the batch SSF with respect to shortening the time requirement and increase in ethanol yield.     

Effect of developmental temperature on swimming performance of zebrafish (<Emphasis Type="BoldItalic">Danio rerio</Emphasis>) juveniles

It is widely known that water temperature affects the swimming capacity of fish. But the effect of the rearing temperature on the swimming ability of the fish at later stages, has not had similar attention. In this study, four populations of zebrafish, were reared in different water temperatures (22, 25, 28 and 31°C) and after being acclimatized in a common temperature (26.5°C) for over a month, they were subjected to swimming trials in order to evaluate the maximum relative critical velocity (RU _crit ) in each case. Fish that were reared in 22°C showed statistically significant lower performance than the ones reared in 31°C (7.72 ± 0.17 vs. 8.79 ± 0.28, means ± S.E.). Possible explanations for the observed differentiation could be the effect of early life temperature on fish muscle ontogeny or on body shape.     

Biodegradation of methyl violet by Pseudomonas mendocina MCM B-402

Pseudomonas mendocina MCM B-402 was found to utilize a triphenylmethane dye, methyl violet as the sole source of carbon when incorporated in synthetic medium. Almost complete decolorization of methyl violet by P. mendocina was observed within 48 h of incubation at ambient temperature (28 ± 2 °C) under aerated culture conditions, when the bacteria were inoculated into Davis Mingioli's synthetic medium at a concentration of 100 mg/l medium. Methyl violet was mineralized to CO₂ through three unknown intermediate metabolites and phenol. The decolorization of the dye involved demethylation. Received: 27 November 1998 / Received revision: 2 March 1999 / Accepted: 5 March 1999     

Adaptation and acclimation of growth and photosynthesis of five Antarctic red algae to low temperatures

Temperature requirements for growth, photosynthesis and dark respiration were determined for five Antarctic red algal species. After acclimation, the stenothermal species Gigartina skottsbergii and Ballia callitricha grew at 0 or up to 5 °C, respectively; the eurythermal species Kallymenia antarctica, Gymnogongrus antarcticus and Phyllophora ahnfeltioides grew up to 10 °C. The temperature optima of photosynthesis were between 10 and 15 °C in the stenothermal species and between 15 and 25 °C in the eurythermal species, irrespective of the growth temperature. This shows that the temperature optima for photosynthesis are located well below the optima from species of other biogeographical regions, even from the Arctic. Respiratory rates rose with increasing temperatures. In contrast to photosynthesis, no temperature optimum was evident between 0 and 25 °C. Partial acclimation of photosynthetic capacity to growth temperature was found in two species. B. callitricha and Gymnogongrus antarcticus acclimate to 0 °C, and 5 and 0 °C, respectively. But acclimation did in no case lead to an overall shift in the temperature optimum of photosynthesis. B. callitricha and Gymnogongrus antarcticus showed acclimation of respiration to 5 °C, and P. ahnfeltioides to 5 and 10 °C, resulting in a temperature independence of respiration when measured at growth temperature. With respect to the acclimation potential of the species, no distinction can be made between the stenothermal versus the eurythermal group. (Net)photosynthetic capacity:respiration (P:R) ratios showed in all species highest values at 0 °C and decreased continuously to values lower than 1.0 at 25 °C. In turn, the low P:R ratios at higher temperatures are assumed to determine the upper temperature growth limit of the studied species. Estimated daily carbon balance reached values between 4.1 and 30.7 mg C g⁻¹ FW day⁻¹ at 0 °C, 16:8 h light/dark cycle, 12–40 μmol m⁻² s⁻¹. Received: 4 November 1999 / Accepted: 7 March 2000     

Effect of seawater temperature on the productivity of <Emphasis Type="Italic">Laminaria japonica</Emphasis> in the Uwa Sea,southern Japan

Recent studies on global climate change report that increase in seawater temperature leads to coastal ecosystem change, including coral bleaching in the tropic. In order to assess the effect of increased seawater temperature on a temperate coastal ecosystem, we studied the inter-annual variation in productivity of Laminaria japonica using long-term oceanographic observations for the Uwa Sea, southern Japan. The annual productivity estimates for L. japonica were 2.7 ± 2.5 (mean ± SD) kg wet wt. m⁻¹ (length of rope) (2003/2004), 1.0 ± 0.6 kg wet wt. m⁻¹ (2004/2005) and 12.1 ± 12.5 kg wet wt. m⁻¹ (2005/2006). Our previous study using the same methodology at the same locality reported that the productivity was estimated for the 2001/2002 (33.3 ± 15.2 kg wet wt. m⁻¹) and 2002/2003 (34.0 ± 8.7 kg wet wt. m⁻¹) seasons. Productivity in 2003/2004 and 2004/2005 was significantly lower than in years 2001/2002, 2002/2003 and 2005/2006. A comparison of oceanographic conditions among the 5 years revealed the presence of threshold seawater temperature effects. When the average seawater temperature during the first 45 days of each experiment exceeded 15.5°C, productivity was reduced to about 10 % of that in cooler years. Moreover the analysis of growth and erosion rates indicates that when the seawater temperature was over 17.5°C, erosion rate exceeded growth rate. Thus, an increase of seawater temperature of just 1°C during winter drastically reduces the productivity of L. japonica in the Uwa Sea.     

Heart rate during development in the turtle embryo: effect of temperature

 Growth and development can occur over a wide range of physical conditions in reptiles. Cardiovascular function must be critical to this ability. However, information on cardiovascular function in developing reptiles is lacking. Previous work indicated that in reptiles the effects of temperature on growth and metabolism are largely restricted to early development. This study examined whether the previously observed effects of temperature and different perinatal patterns of metabolism observed in amniotic vertebrates are correlated with cardiovascular function. Embryonic and hatchling carcass mass, heart mass and heart rate (HR) were compared for snapping turtle eggs (Chelydra serpentina) incubated at 24 ° and 29 °C. Incubation time was shorter at 29 °C (56.2 days) than at 24 °C (71.1 days). Carcass and heart growth showed a sigmoidal pattern at both temperatures. However, cardiac growth showed a relative decrease as incubation proceeded. Incubation temperature significantly affected the HR pattern during development. The HR of embryos incubated at 24 °C was constant for most of incubation (51.8±4.8 min^-1). A small decrease was observed just prior to and a large decrease immediately following hatching (posthatch, 22.3±4.1 min^-1). At 29 °C embryonic HR was greater than at 24 °C early in development (72.3±3 min^-1). The HR steadily decreased to values equivalent to those at 24 °C. The HRs of 24 °C and 29 °C hatchlings were not different. Cardiac output (estimated as the product of heart mass and HR) increased rapidly during early development and then slowed dramatically at both temperatures. These data are consistent with the suggestion that temperature exerts its effects primarily early in development. Furthermore, the changes in cardiovascular function are correlated with metabolic changes in hatching vertebrates. Accepted:12 June 1996     

Isolation and characterization of photoactive complexes of NADPH:protochlorophyllide oxidoreductase from wheat

A photoactive substrate-enzyme complex of the NADPH:protochlorophyllide oxidoreductase (POR; EC 1. 3. 1. 33) was purified from etiolated Triticum aestivum L. by gel chromatography after solubilization of prolamellar bodies by dodecyl-maltoside. Irradiation by a 1-ms flash induced the phototransformation of protocholorophyllide a (Pchlide) with −196 °C absorbance and emission maxima at 640 and 643 nm, respectively. The apparent molecular weight of this complex was 112 ± 24 kDa, which indicates aggregation of enzyme subunits. By lowering the detergent concentration in the elution buffer, a 1080 ± 250-kDa particle was obtained which displayed the spectral properties of the predominant form of photoactive Pchlide in vivo (−196 °C absorbance and fluorescence maxima at 650 and 653 nm). In this complex, POR was the dominant polypeptide. Gel chromatography in the same conditions of an irradiated sample of solubilized prolamellar bodies indicated rapid disaggregation of the complex after Pchlide phototransformation. High performance liquid chromatographic analysis of the POR complexes obtained using two detergent concentrations indicates a possible association of zeaxanthin and violaxanthin with the photoactive complex. Received: 25 February 1998 / Accepted: 8 June 1998     

Enhancement of polysaccharides production in <Emphasis Type="Italic">Ganoderma lucidum</Emphasis> by the addition of ethyl acetate extracts from <Emphasis Type="Italic">Eupolyphaga sinensis</Emphasis> and <Emphasis Type="Italic">Catharsius molossus</Emphasis>

To screen stimulators from Chinese medicinal insects for mycelial growth and polysaccharides production of Ganoderma lucidum, G. lucidum was inoculated into the media with and without supplementation of medicinal insect extracts. The ethyl acetate extract of Eupolyphaga sinensis at 55 mg l⁻¹ lead to significant increase in both biomass and intracellular polysaccharides (IPS) concentration from 8.53 ± 0.41 to 14.16 ± 0.43 and 1.28 ± 0.09 to 2.13 ± 0.11 g l⁻¹, respectively. In addition, the ethyl acetate extract of Catharsius molossus at 55 mg l⁻¹ significantly enhanced extracellular polysaccharides (EPS) production; the EPS yield increased from 350.9 ± 14.1 to 475.1 ± 15.3 mg l⁻¹. There were no new components in the two types of polysaccharides obtained by the addition of the insect extracts.     

Influence of the Selected Antioxidants on the Stability of the Celsior Solution Used for Perfusion and Organ Preservation Purposes

The purpose of the following research was to improve the original Celsior solution in order to obtain a higher degree of stability and effectiveness. The solution was modified by the addition of selected antioxidants such as vitamin C, cysteine, and fumaric acid in the following concentrations: 0.1, 0.3, and 0.5 mmol/l. The solution’s stability was estimated using an accelerated stability test based on changes in histidine concentrations in the solution using Pauly’s method for determining concentrations. Elevated temperatures, the factor accelerating substances’ decomposition reaction rate, were used in the tests. The research was conducted at four temperatures at intervals of 10°C: 60 ± 0.2°C, 70 ± 0.2°C, 80 ± 0.2°C, and 90 ± 0.2°C. It was stated that the studied substances’ decomposition occurred in accordance with the equation for first-order reactions. The function of the logarithmic concentration (log%C) over time was revealed to be rectilinear. This dependence was used to determine the kinetics of decomposition reaction rate parameters (the rate constant of decomposition k, activation energy E _a, and frequency factor A). On the basis of these parameters, the stability of the modified solution was estimated at +5°C. The results obtained show that the proposed antioxidants have a significant effect on lengthening the Celsior solution’s stability. The best results were reached when combining two antioxidants: vitamin C and cysteine in 0.5 mmol/l concentrations. As a result, the Celsior solution’s stability was lengthened from 22 to 299 days, which is 13.5 times. Vitamin C at a concentration of 0.5 mmol/l increased the solution’s stability by 5.2 times (t ₉₀ = 115 days), cysteine at a concentration of 0.5 mmol/l caused a 4.4 times stability increase (t ₉₀ = 96 days), and fumaric acid at a concentration of 0.5 mmol/l extended the stability by 2.1 times (t ₉₀ = 48 days) in relation to the original solution.     

How bees detect coloured targets using different regions of their compound eyes

Honeybees Apis mellifera detect coloured targets presented to the frontal region of their compound eyes using their colour vision system at larger visual angles (α > 15°), and an achromatic visual system based on the long-wave photoreceptor type at smaller visual angles (5° < α < 15°). Here we examine the capability of the dorsal, ventral and frontal regions of the eye for colour detection. The minimum visual angle α_min at which the bees detect a stimulus providing both chromatic contrast and receptor-specific contrasts to the three receptor types varies for the different regions of the eye: 7.1 ± 0.5° for the ventral region, 8.2 ± 0.6° for the dorsal region and 4.0 ± 0.5° for the frontal region. Flight trajectories show that when the target was presented in the horizontal plane, bees used only the ventral region of their eyes to make their choices. When the targets appeared dorsally, bees used the frontodorsal region. This finding suggests that pure dorsal detection of coloured targets is difficult in this context. Furthermore, α_min in the ventral plane depends on receptor-specific contrasts. The absence of S-receptor contrast does not affect the performance (α_min = 5.9 ± 0.5°), whilst the absence of M- and L-receptor contrast significantly impairs the detection task. Minimal visual angles of 10.3 ± 0.9° and 17.6 ± 3°, respectively, are obtained in these cases. Thus, as for many visual tasks, the compound eye of the honeybee shows a regionalisation of colour detection that might be related to peripheral or central specialisations. Accepted: 28 September 1999